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Male Breeding Soundness Exam
We offer comprehensive breeding soundness examinations for your stud dogs. This includes a complete physical examination with special attention to the testicles, penis, and prostate, as well as a complete semen evaluation. We review your male's prior breeding history, including litters produced and bitches that missed. Additional testing may include baseline labwork, prostatic and testicular ultrasounds, cultures, and genetic testing.
For questions related to breeding soundness exams, please speak to one of our reproduction technicians – Linn, Kirsten, or Scott.
Semen Collection and Evaluation
The manual collection of semen is a vital component of evaluating your stud dog. The libido of your dog and the quality of the semen sample is improved by the presence of a teaser bitch. You may bring your own teaser or we can provide one for him. We separately collect the different fractions of the ejaculate and utilize a computer-assisted sperm analysis (CASA) machine for thorough evaluation. Our phase contrast microscope and CASA software is state-of-the-art and incredibly precise, providing you with the most detailed information. You will receive a detailed report of your stud dog’s semen analysis.
Your male's semen quality should be evaluated in the month prior to an expected breeding. If he is used frequently, he should be manually collected and evaluated every six months, or any time a breeding does not take.
Options for Semen
Fresh semen | We collect your male, add semen extenders to enhance the motility and lifespan of the ejaculate, and use the sample for breeding immediately with a bitch. |
Fresh-chilled semen | This method of collecting semen, adding extender, slowly cooling, and packaging the specimen for shipment has made long distance breeding possible. The stud dog is manually collected at our office when the owner of the bitch notifies you that their female has reached her fertile period. With fresh-chilled semen, it is the semen, not the bitch, that does the traveling. We can arrange same-day or overnight shipment within the continental United States Monday through Saturday. We recommend that stud dog owners have their dog's semen checked for its ability to withstand the chilling process. Please see semen shipping (below) for more information. |
Frozen | The purpose of freezing your dog's semen is to ensure his breeding availability. Frozen semen is used when a dog is no longer fertile, deceased, or unavailable due to a scheduling conflict. Please see semen cryopreservation (freezing) below for more information. |
Semen Cryopreservation (Freezing)
Our hospital began working with International Canine Genetics in the early 1990s. We are a licensed freezing center and offer semen freezing once a month. Special arrangements can be made to mitigate scheduling conflicts.
Consider freezing your dog's semen to:
- Preserve his genetics for your own future breeding program
- Provide stud service dog sperm bank near me your dog is not available (show/trial schedules, injury, illness)
- Use as back-up for breeding involving winter shipping or difficult travel
- Use for international breeding (Please contact us in advance for individual export regulations.)
The ideal time to collect your dog's semen for freezing is when he is between two and five years of age. The semen of older dogs may be successfully frozen, but often it is more sensitive to the extreme temperature changes in the process. We have successfully produced hundreds of puppies from frozen semen, many from semen frozen 20 or more years ago!!
How do I schedule an appointment?
Our freeze dates are typically the first Friday of every month with Dr. Gatlin. Please contact us below for further details.
- You may contact us at 508-875-7086 to speak with one of our reproductive technicians – Mary, Linn, Kirsten, or Scott. You may also schedule this appointment by contacting [email protected]
- Once your appointment is scheduled, please remember to bring the following paperwork:
- AKC registration
- a copy of current rabies vaccine certificate
- a negative brucellosis test (if not done here).
- Please arrive 15-20 minutes in advance to complete paperwork and run a brucellosis test, if necessary.
How do you know if the semen is good enough to freeze?
Immediately after collection, the semen is fully evaluated. We tell you if the concentration and motility are sufficient to continue the freezing process. Good quality semen at the time of collection is essential to ensure good semen at the time of thaw and insemination. The sample is then extended with buffers that protect the sperm cells during the controlled temperature drop over four hours to -196 C. The semen is packaged into straws. The number of straws to be stored is determined by the initial sperm count. One frozen straw from your dog’s collection is thawed and examined for post-thaw motility and quality.
If your dog has not been collected in the last 6 to 12 months, we do recommend a collection and evaluation appointment prior to the freeze date to assess the quality.
When do I know the results?
At the end of the day, we will call to inform you the number of straws stored, the post-thaw motility, and the number of breedings available from the collection. We will also email you a detailed copy of the final report.
Where is the semen stored?
After your dog’s semen is collected and frozen, it safely stored here until it is shipped to Kansas City, MO, where Zoetis Inc. maintains the largest canine semen storage bank in the world. We will help establish an account for your stud dog through Zoetis’s online portal system. You will be able to access all of your frozen semen information at www.mysecurelineage.com. Once this account is established, Zoetis will contact you regarding pricing and long-term storage information. When your semen is needed, you call them to arrange for your stored semen to be shipped to the inseminating veterinarian.
The method of collecting semen, adding extender, slowly cooling, and packaging the specimen for shipment has made long distance breeding possible. We ask that you arrange for same-day or overnight semen shipment within the continental United States Monday through Saturday.
If your stud’s semen has not been chilled before, we do recommend a pre-shipment evaluation and chill check test, although this is not required. The "chill check" is done by collecting the dog's semen and putting it through the same chilling process as if it was being shipped. We evaluate its quality every 12 hours for 48 hours. This process enables us to determine if overnight shipping is adequate or if same-day service is required.
Please fill out our chilled semen shipment form so we have all the information in advance. You may also contact our office to speak with one of our reproductive technicians.
International semen shipments require significant planning due to strict regulations. It is the client’s responsibility to research specific destination country regulations. Please contact a staff member to discuss further. You and our reproductive liaisons will need to work together to make certain that all of the regulatory requirements are met for international shipments.
Canine brucellosis is a highly contagious infection caused by a bacteria, Brucella canis. Infected dogs develop problems including infertility, spontaneous abortions, stillborn puppies, and systemic illness. Treatment is incredibly difficult and often results in euthanasia. This disease is also zoonotic, which means it can also be spread to people. Due to these concerns, we require negative brucellosis testing on all dogs and bitches every 3 months. This test can be run by most veterinary laboratories, although we do offer in-house testing, if necessary.
Kirsten evaluating semen.
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Animal Breeding and Reproductive Services
in Frederick County
Consistent winner of Frederick's Best Vet, 2007-2021!
Buckeystown Veterinary Hospital offers reproductive services for both companion and farm animal species.
The following reproductive services are offered for the species listed below. If you require a breeding or reproductive service and do not see it listed, contact us for a consultation at:
Breeding & Reproduction Services for Dogs
- Pre-breeding consultation and evaluation
- Brucellosis testing
- Progesterone testing
- Vaginal cytology
- Breeding management
- Artificial insemination
- Surgical insemination
- Pregnancy diagnosis by ultrasound
- “Puppy count” X-rays
- Dystocia management
- Caesarian section
- Neonatal care
- Semen collection and evaluation
- Semen collection and shipment
- Semen collection, freezing, and storage
Breeding & Reproductive Services for Horses
- Pre-breeding consultation and examination
- Breeding soundness examination
- Breeding management
- Artificial insemination using fresh, chilled, or frozen semen
- Pregnancy diagnosis by ultrasound
- Twin management
- Late-term mare evaluation
- Neonatal care
Breeding & Reproductive Services for Cattle
- Breeding management
- Pregnancy diagnosis by palpation or blood test
- Neonatal care
Breeding & Reproductive Services for Small Ruminants
- Breeding management
- Pregnancy diagnosis by ultrasound
- Neonatal care
Breeding & Reproductive Services for Llamas & Alpacas
- Breeding management
- Evaluation of problem breeders
- Pregnancy diagnosis by how to use bank of america mobile app and ultrasound
- Neonatal care
Services We Offer
Male Canine Reproductive Services
If you're ready to breed your stud dog now, or are concerned about the quality of the dog's semen, or if you're just looking to preserve your dog's breeding ability long after he's gone, Innovative Canine Reproduction has solutions for you.
Collection & Evaluation
Most male dogs are easily collected in the office. Semen quality and quantity is assessed.
Frozen Semen, Preparation, Storing and Shipping
Frozen semen technology allows for the infinite preservation of semen. This allows a male's reproductive capacity to outlive him. Additionally, a male’s reproductive capacity can be owned by multiple breeders. It can be dispersed to multiple locations awaiting use including being shipped over international borders when shipping a dog is not practical. Innovative Canine Reproduction stores semen at one of the largest canine sperm banks in the country.
Frozen semen is shipped in dry shippers cooled by liquid nitrogen. This is a safe and effective means of transport. It is shipped overnight via Federal Express domestically. International shipment times vary.
Infertility
There are many causes of infertility in male dogs. The older a dog is and the more he is used, the greater the likelihood of infertility. One of our areas of interest is identifying and addressing the causes of infertility. Since the restoration of fertility is often difficult, storing semen early in a stud dogs life is a wise investment for any breeding program.
Female Canine Reproductive Services
When a bitch comes into season, timing is critical in every step of the breeding process -- from pinpointing ovulation through progesterone testing to selecting the timing of insemination. Innovative Canine Reproduction provides all these services so responsible breeders can maximize the dog's chances at successful pregnancy.
Breeding Timing by Progesterone
Because of the expense, and critical nature of the appropriate time of insemination, progesterone testing is critical to the breeding process. ICR provides accurate progesterone testing and can pinpoint the most optimal day(s) to begin breeding.
Successful breeding depends on determining the optimum time for insemination. Factors that need consideration include how the semen is prepared, how a particular female cycles and her reproductive history. We maintain equipment dedicated and calibrated for progesterone testing, this allows us to provide accurate same day results Monday through Saturday.
Artificial Insemination
Artificial Insemination (AI) techniques include vaginal, trans-cervical and surgical insemination. The best method for a particular breeding is determined by consultation.
Vaginal insemination is the simplest. It can be done by breeders, owners and veterinarians with success. The technique has application for neat (side by side) and for chilled semen.
Using trans-cervical insemination, the semen is deposited into the uterus through the cervical canal. Two techniques are used to accomplish this procedure. At Innovative Canine Reproduction most trans-cervical inseminations are done under sedation using the Norwegian pipette, a specially designed tool developed in Scandanavia and used widely throughout Europe. Dr. Mantell is one of a small number of people in North America trained in this technique. It is a safe and effective method of intra-uterine insemination. The second technique uses optical instruments to visualize the cervix. We use this technique when body conformation precludes use of the Norwegian pipette. The most appropriate technique for an individual is best determined by consultation.
Surgical insemination is the gold standard for artificial insemination. It is the only technique that allows direct examination of the uterus and ovaries if needed. Surgical insemination bypasses any problem related to the condition of the vagina and cervix. It requires general anesthesia and abdominal surgery. It has broad application for canine reproduction in that veterinarians are universally familiar with similar abdominal surgeries.
The most appropriate technique for an individual dog is best determined by consultation.
Whelping Management & C-sections
Care of the whelping female must be prepared for locally. We are happy to consult with your veterinarian bryan adams pnc bank arts center our expertise is needed. Arrangements to care for our local breeders are made individually.
Services
Epididymal Harvesting
Epididymal Sperm Harvesting is a new and unique process in which we can extract semen from the testes after a neuter or after an unforeseen circumstance. Upon extraction, we evaluate, freeze, and store. This is an exciting opportunity for those who wish to preserve their breeding program. We work with your veterinarian and keep all services confidential.
International Canine Semen Bank Atlanta offers an extensive range of professional products and services that will compliment your clients’ wants and needs. ICSB Atlanta is a specialized Canine Semen Bank that offers canine semen cryopreservation services in a mobile and laboratory setting. We also offer multiple dog accounts.
Evaluation and Freezing of Canine Semen
Many working parts go into the evaluation and freezing of canine semen. Upon collection of semen, we evaluate the sperm count and motility to check the quality of the ejaculate. If the sperm meets the criteria, we begin the freezing process.
Fresh and Chilled Shipment of Semen
We take care of the shipment and write an evaluation report to send with the semen from our in-office laboratory. We provide all necessary forms and information needed to successfully ship the semen domestically or out of the country. Our staff is experienced in shipments, allowing us to stay up to date on requirements so your shipment doesn’t run into any unexpected issues with customs.

We offer the following services in our office and mobile laboratory:
Canine semen collection, evaluation, freezing, and storage
DNA typing and identification
Breeding assistance with fresh and cooled semen
Artificial insemination, products, and instructions
Surgical implant referrals
Reproduction workups for studs and bitches
Breeding consultations
Export bloodwork panels for semen export to foreign countries
Fresh and cooled extended semen kits for transportation to the bitch owner
Organize breeding arrangements for bitches including progesterone’s and timing
Freezing Techniques
There are currently two techniques used to freeze canine semen: the straw method and the pellet method. The first successful insemination with frozen canine semen was in 1970, using the pellet method. When freezing canine semen, the pellet method allows changes in temperature to occur very quickly as the pellets are only 1/16 the volume of the .5mL straws. This increases the sperm viability. The American Kennel Club supported an eight-year study (1971-1979) and they concluded that the pellet method has a higher number of viable sperm cells during the freezing process. After thawing the semen, more sperm cells were recovered with a high motility. When comparing the straw method and pellet method, the pelletized canine semen was 20-60% higher depending on the size of the straws that were used. Conception rates of 73-85% are reported by users of the straw method compared to the 93% conception rate when using pellets surgically with proper progesterone tests on the bitch. The litter size using the pellet method averages a normal size while the straw method will often produce smaller than normal litters.
Shipment Instructions
Chilled Semen – Collection and Shipment
- Notify of collection and shipment as soon as possible. This allows us to collect information such as progesterone numbers and female’s progressions.
- Fill out Shipping Chilled Semen form.
- Pay for shipment.
- Collection appointments should be scheduled in the morning for shipping purposes. If a collection must be done after 2:00 pm, it is required for the package to be manually delivered to Fed-Ex.
Frozen Semen – Preparation and Shipment
- Notify ICSB of frozen shipment at least 48-72 hours before shipment departure to allow time for retrieval and preparation of semen and completion of all forms.
Note: Extra fees apply if notified less than 48 hours before shipment departure. - The tank should be shipped early to avoid shipment delays. It should be verified that the facility has an appropriate means for storage.
- Complete all forms and gather all necessary information needed for shipping internationally. Contact the embassy and review additional import/export policies here.
- Fill out Shipping Frozen Semen and Release of Frozen Semen forms. These must be in possession before shipment is sent.
- Pay for the shipment before departure.

Independent and combined effects of diethylhexyl phthalate and polychlorinated biphenyl 153 on sperm quality in the human and dog
Abstract
A temporal decline in human and dog sperm quality is thought to reflect a common environmental aetiology. This may reflect direct effects of seminal chemicals on sperm function and quality. Here we report the effects of diethylhexyl phthalate (DEHP) and polychlorinated biphenyl 153 (PCB153) on DNA fragmentation and motility in human and dog sperm. Human and dog semen was collected from registered donors (n = 9) and from stud dogs (n = 11) and incubated with PCB153 and DEHP, independently and combined, at 0x, 2x, 10x and 100x dog testis concentrations. A total of 16 treatments reflected a 4 × 4 factorial experimental design. Although exposure to DEHP and/or PCB153 alone increased DNA fragmentation and decreased motility, the scale of dose-related effects varied with the presence and relative concentrations of each chemical (DEHP.PCB interaction for: DNA fragmentation; human p < 0.001, dog p < 0.001; Motility; human p < 0.001, dog p < 0.05). In both human and dog sperm, progressive motility negatively correlated with DNA fragmentation regardless of chemical presence (Human: P < 0.0001, r = −0.36; dog P < 0.0001, r = −0.29). We conclude that DEHP and PCB153, at known tissue concentrations, induce similar effects on human and dog sperm supporting the contention of the dog as a sentinel species for human exposure.
Introduction
Over the last four decades, there has been increasing concern over declining human male reproductive health. Reduced sperm counts have been widely used as an index of male subfertility and meta-analytical studies indicate a 50% global reduction in quality from 1938 to 20111,2,3. Sperm morphology has also been reported to decrease over a period of 17 years in France with some geographical regional variation; Aquitaine and Midi-Pyrenees having the lowest morphology combined with the lowest concentrations4,5. These data are indicative of an environmental aetiology and, in support of this, epidemiological studies showing increased incidences of testicular cancer and malformations at birth have been linked to regions with reduced sperm counts6,7.
Temporal trends in human semen quality are paralleled by a similar trend in dogs that live in the human household, where sperm motility declined by 30% over a 26 year period8. In this latter study, all data was generated from a single laboratory using consistent techniques and thus did not suffer from changes in methodology and quality assurance over the time span encompassed in human meta-analytical studies9. These observations support the hypothesis that temporal trends in semen quality, both in the human and dog, are due dog sperm bank near me shared environmental factors and that the dog may be a sentinel for human exposure to such factors. Access to a controlled breeding population of assistance dogs that are routinely sampled for sperm quality provides a cost-effective means of sperm analyses without the stigma and social complications that accompany analogous human studies. Furthermore, there is considerable potential to extend these analyses to any individual or population of dogs. For example, although not investigated in the current study, this could be achieved by semen collection from the tail of the epididymis10 immediately after removal of dog testes at routine surgical neutering; a procedure that is performed on hundreds of thousands of dogs worldwide each year. In addition, semen collections from live dogs is a procedure that is tolerated by a majority of breeds11 unaccustomed to routine fertility monitoring and can therefore easily be carried out by a trained technician.
Declining sperm quality has been linked with the exposure to persistent anthropogenic chemicals, many of which exhibit endocrine disrupting activity6. Although the mechanisms underlying these putative effects are uncertain, historically, the period of fetal development has been highlighted as being particularly sensitive to chemicals with endocrine disrupting activity12. However, a number of studies have shown that environmental chemicals (ECs) are present in semen in a range of species, including the human, raising the possibility of a direct acute effect of chemicals on sperm13,14,15. In support of this theory, an elevated concentration of seminal bisphenol A (BPA) has been associated with infertility in men15,16 and elevated human seminal phthalate metabolites have been associated with reduced sperm counts17. In a separate study, the phthalates DEHP and di-n-butyl-phthalate (DBP) in human semen were reported to be inversely associated with motility and this was confirmed by the direct application of the same phthalates, at seminal concentrations, to sperm in vitro18. By contrast, PCB congeners 118, 126 and 153 were reported to have no negative effect on human sperm motility in vitro, both individually and in combination19. Similar findings have been reported in the dog where DEHP and PCB153, at concentrations detected in dog semen and testis, exhibited inhibitory and stimulatory effects respectively when tested on sperm motility in vitro8. In the same study, both DEHP and PCB153 were detected in a range of dry and wet dog foods indicative of a dietary source. Both DEHP and PCB153 are widely present in the environment and have been detected in tissues/fluids ranging from human breast milk to ovine liver. DEHP is a widely used plasticizer that leaches out into food and liquids and PCBs are lipophilic, and are therefore present in fatty foods20,21,22. Consequently, exposure occurs largely through the diet and these chemicals are deemed as risk factors for reproductive function23,24,25. In support of this, our own published study has shown that DEHP, PCB153 and other PCB congeners are present within both dry and wet dog food sources8 [DEHP: wet and dry food, 0.37 ± 0.10 and 0.20 ± 0.03 μg/g respectively; ∑PCBs: wet and dry food, 1.35 ± 0.5792 and 0.78 ± 0.223 μg/kg respectively; PCB153: wet and dry food, 0.39 ± 0.193 and 0.22 ± 0.11 μg/kg respectively].
Another parameter of ejaculate quality is the proportion of sperm that exhibit DNA fragmentation26. Environmental chemicals have been shown to induce both human and dog sperm DNA fragmentation8,27 and a commercial mixture of PCBs (Arochlor), administered to rats in vivo and added to sperm in vitro, also increased sperm DNA fragmentation28.
In total, these data suggest that environmental chemicals induce similar acute effects on human and dog sperm in vitro: measurement of sperm motility and DNA fragmentation are tried and tested measures of such chemical effects. Since sperm concentration would not alter during the period of in vitro culture and morphology is confounded by abnormality classification, partly due to swelling that may occur during culture and the generation of artefacts during processing, these parameters were not selected for testing acute chemical effects in vitro29,30. Notwithstanding, testing the effects of individual chemicals on sperm functional parameters does not represent “real-life” exposure to a mixture of chemicals, many of which exhibit synergistic, antagonistic or additive effects. Two chemicals known to be present in dog seminal plasma and testis were therefore selected and their effects tested both independently and in combination, on sperm motility and DNA fragmentation in both the human and dog.
Results
Chemical effects on percentage normal sperm motility in human and dog
The analyses of both the human and the dog sperm motility data found the interaction terms between PCB and DEHP to be significant (human p < 0.001; dog p < 0.05). This indicates that the dose response to either chemical was not independent of the level of the other chemical present. Figure 1 illustrates the effects of PCB153 and DEHP, individually and combined, on dog and human sperm motility. In dog sperm, PCB153 in the absence of DEHP induced a dose-dependent inhibitory effect on sperm motility (Fig. 1ai). In the presence of DEHP at 2x and 10x mean testis concentration (MTC), no dose dependent inhibitory effect of PCB153 was observed. However, reduced motility was observed in the presence of 100x DEHP co-incubated with 2x and 10x PCB153 (Fig. 1aii).
Effect of DEHP and PCB153 on dog and human sperm motility. Chemicals tested individually [ai,bi: PCB only; aii,bii: grey bars: DEHP only] and in combination [aii,bii]. Graphs display fixed concentrations of DEHP with increasing concentrations of PCB153 in dog [ai,aii: p < 0.01] and human [bi,bii: p < 0.001] sperm. Grade a motility: >25 μm/s Error bar = 1 Standard Error of Difference. MTC = Mean testis concentration.
Full size image
In contrast to the dog, neither PCB153 nor DEHP, in the absence of the other chemical, influenced human sperm motility (Fig. 1bi,bii). However, in the presence of 2x DEHP, a dose dependent inhibitory effect in response to PCB153 was observed (Fig. 1bii). The inhibitory effect of PCB153 was broadly maintained in the presence of 10x and 100x DEHP with the exception of 10x PCB153/10x DEHP and dog sperm bank near me PCB153/100x DEHP, where no inhibition was observed (Fig. 1bii).
Chemical effects on sperm DNA integrity in human and dog
For both the human and dog DNA fragmentation data, significant (P < 0.001) PCB.DEHP interaction terms were found. Again, this indicates that the response of DNA integrity to one chemical is not independent of the presence or level of the other chemical. Figure 2 illustrates the effects of PCB153 and DEHP, individually and combined, on both dog and human sperm DNA fragmentation using the sperm chromatin dispersion assay. In the dog, PCB153 in the absence of DEHP induced a dose-dependent increase in sperm DNA fragmentation (Fig. 2ai). A similar dose-dependent increase in DNA fragmentation was observed in response to DEHP in the absence of PCB153 (Fig. 2aii). When PCB153 and DEHP were tested in combination, DNA fragmentation was still increased at higher concentrations of PCB153 but the dose-dependent response was blunted (Fig. 2aii). The response of human sperm to PCB153 and DEHP independently and combined, paralleled that observed in the dog. Figure 2b illustrates that human sperm incubated with PCB153 or DEHP in the absence of the other chemical, exhibited a dose-dependent increase in DNA fragmentation (Fig. 2bi,bii). In addition, DNA fragmentation was increased in response to both chemicals tested in combination although additive effects were not apparent.
Effect of DEHP and PCB153 on dog and human sperm DNA fragmentation. Chemicals tested individually [ai,bi: PCB only; aii,bii: grey bars: DEHP only] and in combination [aii,bii]. Graphs display fixed concentrations of DEHP with increasing concentrations of PCB153 in dog [ai,aii: p ≤ 0.001] and human [bi,bii: p ≤ 0.001] sperm. Error bar = 1 Standard Error of difference between means. MTC = Mean testis concentration.
Full size image
Sperm DNA fragmentation and motility correlations
Figure 3 illustrates the relationship between progressive motility and DNA fragmentation in the presence and absence of each chemical independently and combined. Despite chemical effects on sperm motility (Fig. 1) and DNA fragmentation (Fig. 2), the relationship between these two parameters remained the same regardless of the nature of the chemical exposure. All correlations were significant except the human control samples [Dog (Fig. 3i, n = 352): Control; p < 0.05, r = −0.527, n = 22; DEHP; p < 0.05, r = −0.2862, n = 66; PCB-153; p < 0.01, r = −0.3276, n = 66; Mixture; p < 0.0001, r = −0.2826, n = 198; vs Human (Fig. 3ii, n = 288): Control; p > 0.05, r = −0.4374, n = 18; DEHP; p < 0.05, r = −0.3118, n = 54; PCB-153; p < 0.05, r = −0.2994, n = 54; Mixture; p < 0.0001, r = −0.4037, n = 162].
Correlation between progressive motility and DNA fragmentation in both dog and human sperm. Values from 32 sperm assessments (16 treatments, two time points). Each point represents a different sperm culture equating to a total n = 352 [dog] and n = 288 [human]. Colours denote culture media constituents to demonstrate spread: Control (black), DEHP (red), PCB153 (blue) and mixture (green). Dog (i, n = 352): Control; p < 0.05, r = −0.527, n = 22; DEHP; p < 0.05, r = −0.2862, n = 66; PCB153; p < 0.01, r = −0.3276, n = 66; Mixture; p < 0.0001, r = −0.2826, n = 198; vs Human (ii, n = 288): Control; p > 0.05, r = −0.4374, n = 18; DEHP; p < 0.05, r = −0.3118, n = 54; PCB153; p < 0.05, r = −0.2994, n = 54; Mixture; p < 0.0001, r = −0.4037, n = 162]. Confidence bands (95%) plotted for visual purposes only. Progressive motility based on WHO pre-2010 where sperm swimming grades a and b are combined: ≥5 µm/s.
Full size image
Discussion
Data presented in this paper are significant because we conclusively demonstrate that a selected phthalate (DEHP) and PCB congener (PCB153), at concentrations relevant to environmental exposure, reduce dog sperm motility and increase DNA fragmentation in vitro. In the human, our data showing reduced motility and increased DNA fragmentation are indicative of similar sensitivities to these chemicals in both species. Furthermore, when the chemicals were combined and co-incubated with dog or human sperm, the overall impact on motility or DNA fragmentation was dependent on the concentration ratio. This is the first study to select two environmental chemicals and to test them at four concentrations in all possible combinations representative of those found in the male reproductive tract and fluids. In addition, a negative correlation between motility and DNA fragmentation, that incorporates chemical variables, is described in both species.
Our data on the sensitivity of sperm to short term chemical exposure support similar studies using a range of environmentally relevant chemical challenges in vitro. For example, DEHP, DBP and mono-n-butyl phthalate (MBP), are reported to reduce sperm motility in vitro when added at seminal concentrations measured in infertile men18,31. Dog sperm bank near me mixture of PCBs are also reported to reduce sperm motility in both the human and pig32,33 and in the human, p,p’-dichloro-diphenyl-dichloro-ethylene (p,p’-DDE) has been shown to increase Ca2+ uptake by sperm in a mechanism that involves the CatSper channel34.
To minimise the effects of pre-exposure to chemicals present in ejaculates collected for culture, the current study used a concentration range dog sperm bank near me encompassed the reported variability in seminal concentrations and reduced pre-existing chemical concentrations by sperm processing and washing prior to culture. The subsequent comparison to controls, with no further chemical added, provided a means of testing chemical effects. Despite these steps, pre-exposure differences will inevitably exist between, and within, species. For example, in the current study, when DEHP is present at twice the concentration found in testis, dog sperm appears to be less sensitive to lower PCB concentrations than human sperm. In populations of men from Greenland, Sweden, Poland and the Ukraine, increased seminal PCB153 has been consistently associated with reduced sperm motility but not concentration or morphology35,36. In addition, sperm DNA fragmentation was positively associated with PCB153 in European populations but not in samples from Greenlandic men: an observation that likely reflects dog sperm bank near me pre-exposures.
In the current study, our intention was not to measure and equate chemical concentrations in each individual sample with sperm motility and fragmentation, but to use a concentration range previously established in the dog as a ballpark estimate of chemical concentrations in the male reproductive tract8. That is, concentrations found in the dog reproductive tract and seminal plasma, used as an indicator of those likely present in the human. In support of this contention, human seminal PCB (total) and DEHP concentrations reported in populations of fertile men [total PCB: up to 5.8 ng/ml, DEHP: mean of 0.61 μg/ml]37,38 are comparable to those detected in the dog [PCB: 0.26–13.2 ng/ml; DEHP: 0.75–37.5 μg/ml]8.
Notably, elevated concentrations of both chemicals have been linked with reduced human male fertility18. Semen PCB concentrations have been reported to be higher in a population of ‘infertile’ the london west hollywood west hollywood ca (parameters stated as <20 million/ml or <25% progressive motility and/or <30% normal morphology)25. Further, a research group in China reported an association between increased urinary phthalate metabolites, reduced sperm count and increased sperm DNA damage14. Although the mechanism was not proven, the authors suggest that this likely reflects chemical effects on testicular Sertoli and germ cells as reported in animal studies39,40,41,42.
Although both DEHP and PCB153 have been reported to reduce sperm motility and increase sperm DNA fragmentation individually in the human8,18,37, they have not been assessed in combination, at environmentally relevant concentrations, as reported here. A dog sperm bank near me number of studies have investigated some combinations of environmental chemicals for their effect on human sperm function32. Real-life exposure is to a complex mixture of chemicals that are likely to exhibit synergistic or antagonistic, as well as additive effects, on sperm. Mixtures of endocrine disrupting chemicals have been shown to cooperatively increase Ca2+ concentrations in sperm through the activation of the principle calcium channel CatSper43.
In the current study, the mechanisms that underlie the concentration ratio-dependent effects of the two chemicals combined remain uncertain. PCBs and DEHP are considered to be pro-estrogenic and anti-androgenic respectively raising the possibility that a change in concentration ratio may alter the relative activation of sperm estrogen or androgen receptors44,45. Indeed, in a population of ‘infertile’ men (parameters stated as sperm count < 20 × 106, motility < 50%, morphology < 14% normal) androgen receptor expression is reported to positively correlate with sperm motility46 and PCBs are reported to affect sperm concentration and motility relative to the number of CAG repeats in the androgen receptor gene47. This may account for PCB influences on motility even in the presence of DEHP. Estrogenic compounds have also been reported to reduce human sperm motility in vitro via an induction in redox activity and this mechanism has also been linked to the induction of DNA fragmentation by 2-hydroxy estradiol48.
The greater consistency of chemical effects on human and dog sperm DNA fragmentation compared to motility is interesting and emphasises the importance of looking at more than one sperm functional/viability parameter when assessing environmental effects. It is important to note however that despite this subtle difference, the two sperm parameters were highly correlated in both species and remained so in the presence of the chemicals independently and combined. This is an important observation since it has been reported that human male infertility is associated with increased levels of sperm DNA damage and that sperm motility defects are highest in samples with increased DNA fragmentation49,50. This raises the possibility that there may be a similar relationship between sperm DNA fragmentation, motility and fertility in the dog.
In conclusion, we have demonstrated that the use of low dose tissue relevant concentrations of DEHP and PCB153, independently and in combination, negatively impact on sperm motility and DNA fragmentation in samples whitney bank business checking login from humans and dogs. Since these effects are broadly similar in both species, this raises the possibility that the environmental impact of chemicals in the dog may provide a means of investigating pollutant effects on mammalian fertility in a species in which external influences, such as diet, are better controlled than in an equivalent human study.
Methods
Ethical Approval
Human
Semen donations were obtained from anonymous HFEA registered donors (n = 9) attending the fertility unit at Nottingham University Hospitals. Donors provided informed consent for the use of samples in this research project ensuring ‘General Data Protection Regulation’ compliance. Each donor was initially screened following HFEA and British Fertility Society protocols51. No samples from fertility patients were used and all donors completed HFEA consent forms. Ethical approval was obtained from the School of Veterinary Medicine Ethical Review Committee [Reference 1511,150723]. The HFEA consent forms completed by donors and ethical approval documents were also approved by the Chair of the Ethical Review Committee of the Faculty of Medicine and Health Sciences, University of Nottingham. In accordance with the Royal College of Pathologists guidance on the use of pathological specimens [https://www.rcpath.org/resourceLibrary/the-retention-and-storage-of-pathological-records-and-specimens-5th-edition.html], no further ethical approval was required.
Dog
Semen was collected as part of routine reproductive examination of stud dogs subject to owner consent with full GDPR compliance. Due to dog sperm being collected as part of routine reproductive health checks, the economical outlay associated with sample collection was minimised. The dogs resided in the same region of the UK and lived in the modern household with owners briefed on use of controlled diet and exercise regimes. All semen collections were performed in accordance with relevant guidelines and regulations and the collection protocol was approved by the School of Veterinary Medicine Ethical Review Committee (Refs: 208 101012, 513 120117 and 1097 140227).
Human sperm collection and preparation
Samples were liquefied for 20 minutes at room temperature prior to semen preparation. Ejaculate underwent density gradient centrifugation using a Universal 320 R Hettich centrifuge (DJB Labcare, Newport Pagnell, UK) at 25 °C. Briefly, one millilitre of 40% isotonic density gradient was loaded onto one millilitre of 80% isotonic medium [PureSperm 40/80, Nidacon, Sweden]. Two millilitres of liquefied semen were loaded onto both isotonic mediums and reagents centrifuged at 300 × g for 22 minutes. On completion, the sperm rich pellet was re-suspended using 1.5 ml PureSperm wash (pH range of 7.3–8.5; osmolality: 290–300 mOsm/kg H2O, Nidacon, Sweden).
Dog sperm collection
Ejaculate was collected from stud dogs (n = 11) by routine digital manipulation. The sperm rich fraction (fraction 2) was collected into sterile plastic 15 millilitre Greiner centrifuge tubes [Sigma-Aldrich, Dorset, UK]. INRA extending medium [INRA, Nouzilly, France] was added to sperm at a 2:1 ratio to aid sperm survival.
Chemical preparation
The two chemicals selected for co-culture with sperm were diethylhexyl phthalate (DEHP) and polychlorinated biphenyl congener 153 (PCB153) and concentrations were calculated relative to those present in dog testicular tissue8. The rationale for this was (1) dog testis concentrations of both chemicals have been established and standardised in our previous study and are reflective of exposure of the male reproductive tract8 (2) dog and human semen PCB concentrations are variable but generally higher than testis and in range of the concentrations tested in vitro8,25,52 (3) although dog semen DEHP concentrations have not been determined, due to the large amount of dry material required, reported human DEHP concentrations are in range of established dog testis measurements14,18,53. On this basis, dog testis concentrations were used as a standardised measure of environmental exposure relevant to both species. DEHP (CAS no: 117-81-7) and PCB153 (CAS no: 35065-27-1) [Sigma-Aldrich, Dorset, U.K.] were dissolved in 100% dimethylsulphoxide (DMSO) and diluted with PBS to 4x, 20x and 200x mean testis concentration containing 0.02% DMSO. Chemical preparations were co-incubated with sperm at a 1:1 ratio with a final exposure concentration of 2x, 10x and 100x mean testis concentration. Sperm were incubated with each chemical at each concentration individually and with a mixture of the two chemicals in all 16 combination ratios (Table 1). Acute treatment effects were assessed at 10 minutes and 3 hours and control incubations carried out with 0.01% DMSO only.
Full size table
Motility assessment
Sperm dog sperm bank near me was assessed using Computer Assisted Sperm Analysis (CASA) software. Sperm were acclimatised for two minutes on a 37 °C stage prior to assessment. Five µl of sperm, irrespective of species, were pipetted into a specialised 20 µl cellvision glass slide counting chamber (Code CV1020-2CV; CellVision, the Netherlands). A minimum of 200 sperm were assessed for each treatment. For human sperm, motility was tracked by use of the diagnostic software ‘SAMi’ (Procreative Diagnostics Ltd, Staffordshire, UK) and an Olympus BH2 Microscope [KeyMed (Medical and Industrial Equipment) Ltd, Essex, UK]. Dog sperm motility was assessed using the Hobson’s CASA tracking system (Hobson’s tracking systems Ltd., Sheffield, UK) and viewed using a negative-high phase contrast objective (x20) on an Olympus BH2 microscope dog sperm bank near me with a camera ocular. For both species, sperm motility was assessed according to WHO 199954 where grade a motility was classified as ≥ 25 µm/s and progressive motility ≥ 5 µm/s (grades a & b combined).
DNA fragmentation
Sperm DNA fragmentation was quantified by defragmentation index (sDFI %) measured using the sperm chromatin dispersion assay55. Sperm were assessed based on the size of the halos, indicative of sperm that exhibited nuclei with non-fragmented DNA. Sperm with denatured or fragmented DNA were identified by the absence of a halo, or a halo that was exceedingly small. Briefly, equal volumes of intact, unfixed sperm and 1% agarose solution were combined. Fifteen microliter aliquots of sperm-agarose suspensions were pipetted onto agarose pre-coated poly-L-lysine slides (CAS: P4981; ThermoFisher Scientific Ltd. UK), covered with a 22 mm × 22 mm cover-slip, and placed in a 4 °C environment for five minutes to fix the sample. After an acid treatment (0.08 M HCl) of seven minutes, sperm were incubated in a lysing reagent (0.8 M DTT, 0.4 M Tris, 2 M NaCl, 1% triton-X) for 20 minutes. Slides were then submerged in dH2O for a period of five minutes followed by dehydration through a series of ethanol solutions (70%, 90% and 100% respectively). Visualisation was obtained using Diff-Quick staining reagents (eosinophilic and basophilic stains; CAS 9990700, ThermoFisher Scientific Ltd, Paisley, UK) and analysis undertaken using oil immersion, at 1000x magnification [Leica DM 5000 B microscope; Leica Microsystems, Milton Keynes, UK]. A minimum of 200 sperm were assessed for each treatment. Positive controls were incubated with 300 µm H2O2 before denaturation and negative controls by omission of the denaturation step. To determine the defragmentation index, the number of fragmented sperm was divided by the total number of sperm counted. This provided a value for the proportion of sperm that were fragmented. This proportion was then plotted on the logit scale.
Experimental design and Statistical analysis
For both the human and dog experiments, sperm samples were treated with PCB153 at one of four concentrations in combination with DEHP, also at one of four concentrations. The concentrations used for both chemicals corresponded to 0x, 2x, 10x and 100x the baseline concentration measured from dog testes. Thus there were 16 possible combinations of PCB153 and DEHP. Each combination was randomly allocated to one of 16 separate sub-samples of sperm, from each replicate donor.
Statistical analysis was undertaken using GenStat 17th edition (VSN International Ltd, Hempstead, UK). The proportions out of known numbers of sperm that had a particular characteristic were analysed as grouped binary data by fitting a generalised linear mixed model with a logit link function, assuming a binomial error distribution. The fixed effects included in the statistical model were PCB, DEHP and the PCB.DEHP interaction term. Another fixed effect, TIME, and its interaction terms were included when two repeated samples from the same tube were measured three hours apart. The random effects were donor and culture-tube within donor.
The statistical significance of a fixed effect was tested using an F-Ratio test. The analysis output provided predicted mean proportions and standard errors of difference between means which were graphically represented on a logit scale. Logit values were converted back into proportions and graphically plotted [GraphPad Prism 7.0, GraphPad Ltd, California, USA]. A single error bar on each figure represents the standard error of the difference between means. Where correlations were investigated, data was assumed to be non-normally distributed and a Spearman’s rank correlation analysis was undertaken to provide correlation coefficients between motility and DNA fragmentation.
Data Availability
The datasets generated and analysed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
We thank Karen Pooley for the management of the human donors attending the Fertility Unit at Nottingham University Hospital and for expert technical advice and assistance. We also thank Natasha White for the collection of ejaculates from a population of assistance dogs. The work was supported by University of Nottingham internal funds.
Author information
Rebecca N. Sumner
Present address: Hartpury University, Gloucester, GL19 3BE, UK
Affiliations
School of Veterinary Medicine & Science, University of Nottingham, Sutton Bonington, LE12 5RD, UK
Rebecca N. Sumner, Gary C. W. England & Richard G. Lea
Fertility Unit, East Block B floor, Nottingham University Hospital, Nottingham, NG7 2UH, UK
Mathew Tomlinson
School of Biosciences, University of Nottingham, Sutton Bonington, LE12 5RD, UK
Jim Craigon
Contributions
R.G.L. and R.N.S. conceived of and designed the study. M.T. managed the human donors and collection of human semen samples and G.C.W.E. managed the collection of canine samples. All authors provided academic input into the paper. R.N.S. carried out the computer assisted sperm analysis, chromatin dispersion assay and subsequent analysis. J.C. provided expert statistical advice and contributed to the analysis. R.G.L. and R.N.S. wrote the paper.
Corresponding author
Correspondence to Richard G. Lea.
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Sumner, R.N., Tomlinson, M., Craigon, J. et al. Independent and combined effects of diethylhexyl phthalate and polychlorinated biphenyl 153 on sperm quality in the human and dog. Sci Rep9, 3409 (2019). https://doi.org/10.1038/s41598-019-39913-9
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ii)Reduced semen collection frequency – semen freezing provides an alternative/supplement to the demands of natural/chilled semen breeding regimes. With semen freezing the collection frequency can be tailored to the dog's physical needs and semen quality. The bank of semen collected can be used to aid his breeding targets, whether as a complete alternative or supplement to natural/chilled semen collections.
3.More efficient use of your dog’s semen
When breeding via natural methods or chilled AI, the dog’s ejaculate is dedicated to one bitch or, in the case of chilled semen, is divided between bitches according to demand with unused semen discarded after 24 hours. With frozen semen, some dogs can split one ejaculate into multiple breeding doses which can be frozen and kept indefinitely, preventing semen wastage and permitting more efficient use of your stud dog.
4. Logistics and a worldwide market
i)A further advantage of frozen semen is the ability to ship the semen ahead of the bitch’s planned time of requirement. Most vet practices now have frozen semen storage, enabling bitch owners to order semen weeks, if not months, in advance and so prevent the stress to all parties involved in timing semen arrival against the bitch’s cycle.
ii)The indefinite shelf life of frozen semen providing its maintenance at -196 degrees Celsius also means it can be shipped worldwide where regulations and transport permits. This opens up the possibility of an exciting worldwide market for you and your dog – the world becomes your dog's oyster with frozen semen distribution.
The Service
Our semen collection and freezing service includes;
Semen collection by our experienced team members
Processing & freezing into 0.5ml straws in our state of the art laboratory
Full post thaw analysis & quality control written report
Once the semen is frozen it is transferred into our permanent storage facility, see ‘Frozen Semen Storage & Distribution’ for more details on storage.
How many doses will my dog produce from one collection?
The number of doses that each dog produces is highly variable, the table below shows the estimated doses calculated using body weight. It may be possible to carry out a supplementary collection on the same day to increase sample size, allowing for greater straw numbers.
Small Dog | Up to 10Kgs | 1+ Doses |
Medium Dog | Up to 30Kgs | 2+ Doses |
Large Dog | Up to 40Kgs | 3+ Doses |
Extra Large Dog | 40Kgs + | 4+ Doses |
This Months Deals!
We run regular semen freezing clinics where you can save over £40 on frozen semen collections. To find out when our next date is click here.
Have you joined our loyalty scheme yet? click here to claim your FREE semen freeze
Please contact us for more information and click here to book.
More Info
We are proud to be an AKC approved Freezing Center. Here are some of the frequently asked questions regarding canine semen freezing:
1) Why should I freeze my dog’s Semen?
Long Term Storage: To ensure breeding availability for future generations, you should have his semen frozen if your stud has qualities which are valuable.
For Breeding When the Stud is Not Available: Live breeding can be limited by the stud’s show or trial schedule, overbooking for the stud’s service or other scheduling conflicts. The use of frozen semen allows availability during the bitch’s fertile time.
Long-Distance and International Breeding: Long-distance breedings may be accomplished using semen which has been either fresh-extended or frozen and eliminates the need to transport the bitch or stud dog.
2) How is Semen collected?
Semen is collected from the stud dog by manual stimulation; the different parts or fractions of the ejaculation are collected separately, so that good quality sperm-rich semen is frozen and stored. In general, semen of better quality with higher sperm count is collected when the dog’s libido is high. Therefore, try to closely approximate a typical breeding situation for each stud: owners are encouraged to provide a bitch in season to use as a “teaser”. In addition, if the dog associates a particular item with breeding, such as a rug, table, breeding rack, etc., that item should be brought to the collections.
3) What kind of paper work is needed to collect my stud?
- A copy of the stud’s individual registration papers.
- Positive identification such as a tattoo or microchip.
- Completed Semen Freeze Agreement which can be emailed to you.
- Completed Semen Freeze Authorization which can be emailed to you.
- A copy of the DNA Profile for your dog (which is required by AKC). If you do not have a DNA Profile on your dog, we will submit a cheek swab sample to the AKC.
- A copy of a negative Brucellosis test. If your stud has not had a Brucellosis test performed then we can have the test performed the same day as the collect and freeze. This test is a MUST in order for your males semen to be stored in our liquid nitrogen tanks.
4) What does it cost to freeze semen?
Semen freezing is a multi-step process utilizing a variety of equipment and materials. Please see our Reproduction price sheet in the forms section of our website for detailed information.
5) How do I set up an appointment for Semen Freezing?
Once you have reviewed the information, please call us at 281-443-2362 or email us at [email protected] to set up an appointment. The receptionist will take your information and our manager Jenny will return your call. She will then discuss the process and set up the appointment with you. We are only able to do collect and freezing on certain days so we suggest you schedule far in advance. Dr. McGuire is our only vet that is able to perform this and she fills up quickly and can only manage at least 1- 2 freezes a day. The collection part is easy it is the freezing that is very long and tedious.
Scheduled freezes that do not show up will not be allowed to schedule again. Also, if your dog is unable to be collected or his semen is not viable and cannot be frozen there is still a charge for time and supplies used which is $150.
Semen collection in the dog
This review will discuss semen collection in the dog. Semen samples may be collected from male dogs for the purposes of artificial insemination, cryopreservation or diagnosis. The materials needed for semen collection depend on which method is used and the collector's level of expertise with this procedure. At minimum, two sterile centrifuge tubes or specimen cups can be used to collect semen as it is ejaculated (for the combined first and second fractions and for the third fraction). The most common method for semen collection in the dog is by digital stimulation. Under ideal conditions, this procedure is performed in the presence of an estrous bitch. Initially, the dog's penis is vigorously massaged through the prepuce at the level of the bulbus glandis (caudal-most aspect of the prepuce) until a partial erection develops (initial engorgement of the bulbus glandis). The prepuce is quickly retracted past the bulbus glandis and firm constant pressure is applied to the penis behind the bulbus glandis by squeezing the penis between index finger and thumb. Pelvic thrusting may occur following application of pressure behind the bulbus glandis during the development a "full" erection. The ejaculate is composed of three fractions: first (sperm-poor), second (sperm-rich) and third (prostatic fluid). In addition to digital stimulation of the penis, spermatozoa have been collected from dogs using electroejaculation and pharmacologic methods.
Services We Offer
Male Canine Reproductive Services
If you're ready to breed your stud dog now, or are concerned about the quality of the dog's semen, or if you're just looking to preserve your dog's breeding ability long after he's gone, Innovative Canine Reproduction has solutions for you.
Collection & Evaluation
Most male dogs are easily collected in the office. Semen quality and quantity is assessed.
Frozen Semen, Preparation, Storing and Shipping
Frozen semen technology allows for the infinite preservation of semen. This allows a male's reproductive capacity to outlive him. Additionally, a male’s reproductive capacity can be owned by multiple breeders. It can be dispersed to multiple locations awaiting use including being shipped over international borders when shipping a dog is not practical. Innovative Canine Reproduction stores semen at one of the largest canine sperm banks in the country.
Frozen semen is shipped in dry shippers cooled by liquid nitrogen. This is a safe and effective means of transport. It is shipped overnight via Federal Express domestically. International shipment times vary.
Infertility
There are many causes of infertility in male dogs. The older a dog is and the more he is used, the greater the likelihood of infertility. One of our areas of interest is identifying and addressing the causes of infertility. Since the restoration of fertility is often difficult, storing semen early in a stud dogs life is a wise investment for any breeding program.
Female Canine Reproductive Services
When a bitch comes into season, timing is critical in every step of the breeding process -- from pinpointing ovulation through progesterone testing to selecting the timing of insemination. Innovative Canine Reproduction provides all these services so responsible breeders can maximize the dog's chances at successful pregnancy.
Breeding Timing by Progesterone
Because of the expense, and critical nature of the appropriate time of insemination, progesterone testing is critical to the breeding process. ICR provides accurate progesterone testing and can pinpoint the most optimal day(s) to begin breeding.
Successful breeding depends on determining the optimum time for insemination. Factors that need consideration include how the semen is prepared, how a particular female cycles and her reproductive history. We maintain equipment dedicated and calibrated for progesterone testing, this allows us to provide accurate same day results Monday through Saturday.
Artificial Insemination
Artificial Insemination (AI) techniques include vaginal, trans-cervical and surgical insemination. The best method for a particular breeding is determined by consultation.
Vaginal insemination is the simplest. It can be done by breeders, owners and veterinarians with success. The technique has application for neat (side by side) and for chilled semen.
Using trans-cervical insemination, the semen is deposited into the uterus through the cervical canal. Two techniques are used to accomplish this procedure. At Innovative Canine Reproduction most trans-cervical inseminations are done under sedation using the Norwegian pipette, a specially designed tool developed in Scandanavia and used widely throughout Europe. Dr. Mantell is one of a small number of people in North America trained in this technique. It is a safe and effective method of intra-uterine insemination. The second technique uses optical instruments to visualize the cervix. We use this technique when body conformation precludes use of the Norwegian pipette. The most appropriate technique for an individual is best determined by consultation.
Surgical insemination is the gold standard for artificial insemination. It is the only technique that allows direct examination of the uterus and ovaries if needed. Surgical insemination bypasses any problem related to the condition of the vagina and cervix. It requires general anesthesia and abdominal surgery. It has broad application for canine reproduction in that veterinarians are universally familiar with similar abdominal surgeries.
The most appropriate technique for an individual dog is best determined by consultation.
Whelping Management & C-sections
Care of the whelping female must be prepared for locally. We are happy to consult with your veterinarian when our expertise is needed. Arrangements to care for our local breeders are made individually.
Basic information about the cryopreservation of canine sperm in Genomia sperm bank
The Genomia sperm bank produces cryopreserved insemination doses (deep frozen).
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Semen collection
The semen collection is the first step of the whole process of production of frozen insemination doses. The semen is collected by veterinarians from the veterinary clinic Vedilab in Pilsen we cooperate with. The sperm collection is made via manual massage that is a painless and pleasant method for most of the dogs. If your dog has never undergone a semen collection, we recommend that a female in oestrus is present during the semen collection; otherwise the collection may not be successful. However, if the dog has experience in semen collection, the presence of an oestrous teaser bitch is not required. The collection which is done prior to the whole process of semen freezing is to be planned at least two weeks in advance via phone communication. Please call us or send us an email message with basic information and your phone number. We will contact you and agree on all details. To increase the yield of the insemination doses the collection is made twice with a short interval between the collections (interval of approx. 30 minutes). And during this waiting time, we can complete together the documents required. (Information for future semen collection by other veterinarian). It is necessary for cryopreservation of the sperm that the dog is not used for reproductive purposes for at least one month or two months when the dog is used for mating more frequently. The ejaculate collection is the only step in the whole process you should attend. The whole „action” will not last more than one hour.
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Transport, analysis, freezing (cryopreservation)
The collected ejaculate is transported to our laboratory, where a detailed analysis of the semen is performed. If the semen meets the minimum requirements for the semen suitable for freezing, the processing is continued. Upon analysis the ejaculate is treated with a special diluent (extender) that protects the semen from damage due to very low temperatures. The diluted ejaculate is filled into thoroughly identified sperm straws where the insemination doses are stored. This step is followed by freezing that is finished by putting the insemination doses into liquid nitrogen where they are stored until the moment you decide to use them. One sperm straw is unfrozen 24 hours after freezing and is used to evaluate whether the cryopreservation was successful. Now, the cryopreservation can be considered successful and you will receive a report of the semen quality and number of the insemination doses stored.
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Usage of cryopreserved semen dose
You can use the insemination doses of your dog at any time or, for example, you can transfer them. The transport of the insemination doses in liquid nitrogen requires a special approach, so inform us please sufficiently in advance.
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Benefits of natural reproduction and insemination
Price list of Genomia sperm bank
